The carbon we do not see—the impact of low molecular weight compounds on carbon dynamics and respiration in forest soils: a review

Dissolved organic matter (DOM), typically quantified as dissolved organic carbon (DOC), has been hypothesized to play many roles in pedogenesis and soil biogeochemical cycles, however, most research to date concerning forest soils has focussed on the high molecular weight (HMW) components of this DOM. This review aims to assess the role of low molecular weight (LMW) DOM compounds in the C dynamics of temperate and boreal forest soils focussing in particular on organic acids, amino acids and sugars. The current knowledge of concentrations, mineralization kinetics and production rates and sources in soil are summarised. We conclude that although these LMW compounds are typically maintained at very low concentrations in the soil solution (<50 μM), the flux through this pool is extremely rapid (mean residence time 1-10 h) due to continued microbial removal. Due to this rapid flux through the soil solution pool and mineralization to CO₂, we calculate that the turnover of these LMW compounds may contribute substantially to the total CO₂ efflux from the soil. Moreover, the production rates of these soluble transitory compounds could exceed HMW DOM production. The possible impact of climate change on the behaviour of LMW compounds in soil is also discussed.     

红壤中低分子量有机酸的吸附动力学

低分子量有机酸在土壤中广泛存在尤其是在根际土壤中,其主要来源于动植物残体的分解、微生物的代谢、植物根系的分泌和土壤中有机物的转化等。由于羧基的强络合作用,低分子量有机酸一旦进入土壤将很快被土壤吸附,影响土壤的表面电荷性质和动电性质,进而影响其在土壤中的作用。关于低     

低分子量有机酸对红壤磷酸单酯酶活性的影响

采用室内模拟试验研究了低分子量有机酸(柠檬酸、草酸、苹果酸、酒石酸)对红壤磷酸单酯酶活性的影响。结果表明,低浓度有机酸(<1μmol g-1)对磷酸单酯酶活性有促进作用,且促进作用大小依次为柠檬酸≈草酸>苹果酸>酒石酸;而高浓度有机酸(>5μmol g-1)则为抑制作用,且抑制作用大小依次为柠檬酸>草酸>苹果酸>酒石酸。当体系pH趋向酶促反应最佳pH时,磷酸单酯酶活性增强;反之,当体系pH远离酶促反应最佳pH时,磷酸单酯酶活性减弱。有机酸根一方面通过羧基的辅助作用提高磷酸单酯酶活性;另一方面通过释放土壤A l3+、Fe3+等金属离子,对土壤磷酸单酯酶活性有一定的抑制作用。     

红壤主要土壤组分对低分子量有机酸吸附的研究

通过一次平衡法研究了双氧水去除有机质、添加1%腐殖酸和DCB法(连二亚硫酸钠-柠檬酸钠-碳酸氢钠)去除游离氧化铁、铝对红黏土发育的红壤吸附草酸、柠檬酸、酒石酸和苹果酸的影响。结果表明,去除有机质后,由于受溶液pH、表面吸附点位变化、土壤结构变化、表面基团活性变化、草酸根生成等多种因素的影响,红壤对低分子量有机酸的吸附量虽略有增加,但增加并不明显。添加腐殖酸培养一个月后,由于受土壤表面电荷变化、吸附点位覆盖、氧化铁活性改变、基团质子化等因素的影响,红壤对低分子量有机酸的吸附量虽有轻微的减少,但减少也不明显。去除占土壤总量3.79%的游离氧化铁、铝后,土壤表面正电荷将会显著减少,而导致红壤对低分子量有机酸的吸附量显著减少,其减少程度因有机酸种类而异,与原土对有机酸的最大吸附量(Sm)大小顺序相反。     

温度和水土比对红壤吸附低分子量有机酸的影响

通过一次平衡法考察了温度和水土比对红壤吸附草酸、柠檬酸、酒石酸和苹果酸的影响。结果表明,有机酸的吸附量、最大吸附量（Sm）及吸附亲和力常数（K）均随温度的升高而增加,随着水土比的增加而降低。温度和水土比变化对有机酸吸附的影响程度因有机酸种类而异,影响程度大小与土壤对有机酸的吸附亲和力（K）大小顺序相反。45℃ 时,土壤对草酸、柠檬酸、酒石酸和苹果酸的吸附量分别是 25℃ 时的 1.19、1.22、1.24 和 1.28 倍;100:1 水土比条件下,草酸、柠檬酸、酒石酸和苹果酸的吸附量分别较 10:1 水土比条件下减少了 48.3%、54.9%、63.5% 和 76.1%。     

pH、离子强度和介电常数对低分子量有机酸在红壤中吸附行为的影响

通过一次平衡法考察了pH、离子强度和溶剂介电常数对红壤吸附低分子量有机酸的影响。研究结果表明,随着溶液pH值的升高有机酸的吸附量降低,其中草酸和酒石酸的吸附量在pH 3.5 ~ 5.0范围内随着pH值的升高而急剧下降,之后缓慢下降（pH 5.0 ~ 7.0）。柠檬酸、酒石酸和苹果酸的吸附量在1 ~ 2 mmol/L初始浓度范围内随着离子强度的增加没有明显变化,但在2 ~ 20 mmol/L初始浓度范围内却随着离子强度的增加而增加。红壤对草酸、柠檬酸、酒石酸和苹果酸的吸附量均随着溶剂介电常数的减小而增加。当溶剂中含6% 的乙醇时,草酸、柠檬酸、酒石酸和苹果酸的吸附量分别是对照（不含乙醇）的1.05、1.05、1.11和1.31倍。     

Root uptake of N-containing and N-free low molecular weight organic substances by maize: A C/N tracer study

Most studies showing potential organic nitrogen uptake were conducted with amino acids. They conclude that, in some ecosystems, amino acids significantly contribute to the N demand of plants and that roots have special transporters to re-uptake amino acids released into the rhizosphere. However, the relevance of the uptake of organic N compounds can only be evaluated by comparing the uptake of N-containing and N-free organic substances. We compared the uptake of alanine, glucose and acetate labelled with ¹⁴C by maize. Additionally, the N uptake was estimated by ¹⁵N labelled alanine and KNO₃. We found a similar uptake of ¹⁴C from alanine, glucose and acetate, amounting for the whole plant less than 1% of ¹⁴C input. These results show that maize did not prefer N-containing to N-free organic substances. The uptake of ¹⁵N by maize exceeded that of ¹⁴C (10- to 50-fold), irrespective of the ¹⁵N source. However, plant uptake of nitrate (23.6–35.2% of ¹⁵N input) always exceeded the uptake of N from alanine (9.6–28.8%). The uptake of organically bound N by maize growing in soil occurred mainly by transpiration flow – as dissolved organics. The contribution of specific amino acid transporters was minor.     

低分子量有机酸对红壤无机磷活化的作用

采用室内模拟试验研究了低分子量有机酸,如草酸、柠檬酸、酒石酸和苹果酸在红壤无机磷活化中的作用。结果表明,相同浓度下,有机酸活化土壤磷的能力为柠檬酸酒石酸苹果酸。低浓度(0.5mmolL-1)时,草酸活化能力最小;高浓度(≥5mmolL-1)时,其活化能力最大。对同一种有机酸而言,土壤各无机磷形态活化量均随pH的升高而降低;且在同一酸度下,其活化量以铝磷(Al-P)为最多,铁磷(Fe-P)和钙磷(Ca-P)次之,闭蓄态磷(O-P)则最少。有机酸活化土壤无机磷酸盐应该是质子酸效应和有机酸阴离子络合效应共同作用的结果,且与磷酸盐的溶度积常数密切相关。研究结果对根系土壤无机磷素循环研究有着重要的意义。     

The effects of Fe deficiency on low molecular weight organic acid exudation and cadmium uptake by Solanum nigrum L.

Abstract

The influence of Fe-deficiency on the root exudation of low molecular weight organic acids (LMWOAs), pH alteration and cadmium (Cd) accumulation and translocation were investigated in morel (Solanum nigrum L.) in hydroponic culture experiments. Tartaric, citric, malic and acetic acids were monitored because these acids were abundant and often detected as root exudates. Results showed that Fe-deficient plants excreted large amounts of LMWOAs in comparison with Fe-sufficient plants across all Cd treatments (p <0.05). In both cases the concentrations of the four organic acids were tartaric > citric > malic > acetic. The results showed that the Fe-deficient plants with higher concentrations of LMWOAs accumulated more Cd (p <0.05) and induced a decrease in solution pH compared with the Fe-sufficient plants. Cadmium accumulation in the Fe-deficient and Fe-sufficient plants had significant positive correlations with the exudation of malic and acetic acids (p <0.05 and p<0.01). Cadmium accumulation in the Fe-sufficient plants had a significant (p<0.01) positive correlation with the exudation of tartaric acid, whereas there was a negative correlation (p<0.01) between Cd accumulation and the exudation of tartaric acid in the Fe-deficient plants. No significant correlation between the exudation of citric acid and Cd accumulation was obtained. Our results indicate that Fe-deficiency could induce Cd accumulation and translocation through an increase of LMWOAs exudation and pH alteration, both of which enhance Cd bioavailability.     

Loss of low molecular weight dissolved organic carbon (DOC) and nitrogen (DON) in H2O and 0.5 M K2SO4 soil extracts

Soil extracts are routinely used to quantify dissolved organic nutrient concentrations in soil. Here we studied the loss and transformation of low molecular weight (LMW) components of DOC (¹⁴C-glucose, 1 and 100 μM) and DON (¹⁴C-amino acid mixture, 1 and 100 μM) during extraction of soil (0-6 h) with either distilled water or 0.5 M K₂SO₄. The extractions were performed at 20 °C, at 4 °C, or in the presence of an inhibitor of microbial activity (HgCl₂ and Na-azide). We showed that both glucose and amino acids became progressively lost from solution with increasing shaking time. The greatest loss was observed in H₂O extracts at 1 μM for both substances (>90% loss after 15 min). Lower temperature (4 °C) and presence of K₂SO₄ both resulted in reduced loss rates. The presence of microbial inhibitors effectively eliminated the loss of glucose and amino acids. We conclude that microbial transformation of LMW-DOC and DON during H₂O or K₂SO₄ extraction of soil may affect the estimation of their concentrations in soil. This finding has significant implications for methods that rely on chemical extractions to estimate LMW-C components of DOC and DON.     

Liquid extraction of low molecular mass organic acids and hydroxamate siderophores from boreal forest soil

Low molecular mass organic acids (LMMOAs) and hydroxamate siderophores (HS) are molecules secreted by microbes and have previously been found in soil solution and in cultures. Mycorrhizal fungi are suggested to be involved in the nutrient uptake processes of trees and weathering of minerals. In this study soil samples taken from the O and E horizons of a podzol were extracted with 10 mM potassium phosphate buffer at pH 7.2. Variable parameters included addition of methanol to the extraction buffer and the use of ultrasonication or rotary shaking during extraction. LMMOAs and HS content of the soil extracts were determined. Analysis of soil extracts were carried out by liquid chromatography mass spectrometry (LC–MS) and the extraction results compared to results for soil solution samples obtained by centrifugation of the soils sampled. The extraction yields were significantly increased by addition of methanol to the extraction buffer, especially for the O horizon samples. Rotary shaking of the samples for 90 min gave slightly higher yields than ultrasonication for 15 min but the reduction in extraction time makes ultrasonication an attractive option. Of the HSs determined, ferricrocin was found in all samples. Optimal extraction conditions showed citric acid and isocitric acid to be the most abundant organic acids in the O and E horizons, respectively.     

Carbon input and crop-related changes in microbial biomarker levels strongly affect the turnover and composition of soil organic carbon

It is increasingly recognized that a detailed understanding of the impacts of land use on soil carbon pools and microbially mediated carbon dynamics is required in order to accurately describe terrestrial carbon budgets and improve soil carbon retention. Toward this understanding, we analyzed the levels of biomarkers including phospholipid fatty acids, amino acids, monosaccharides, amino sugars, and several indicators of labile and stabilized carbon in soil samples from a long-term agricultural field experiment. Our results imply that the composition of soil organic carbon (SOC) strongly depends on both the applied fertilization regime and the cultivated crop. In addition, our approach allowed us to identify possible mechanisms of microbial growth and contributions to soil carbon storage under different long-term agricultural management regimes.Amino acids and monosaccharides were quantitatively the most dominant biomarkers and their levels correlated strongly positively with microbial biomass. The relative contributions of the studied biomarkers to the total SOC varied only slightly among the treatments except in cases of extreme fertilization and without any fertilizer. In case of extreme fertilization and with alfalfa as crop type, we found evidence for accumulation of microbially derived monosaccharides and amino acids within the labile OC pool, probably resulting from soil C saturation. Interestingly, we also found an accumulation of microbially derived monosaccharides and amino acids in completely unfertilized plots, which we assumed to be caused by the smaller pore space volume and subsequent oxygen limitation for microbial growth. Mineral fertilization also had substantial effects on soil organic N when applied to plots containing alfalfa, a leguminous plant. Our results demonstrate that over-fertilization, fertilizer type, and the cultivated crop type can have major impacts on the turnover and composition of soil organic carbon, and should be considered when assessing management effects on soil C dynamics.     

Sorption, microbial uptake and decomposition of acetate in soil: Transformations revealed by position-specific C labeling

Many previous studies on transformation of low molecular weight organic substances (LMWOS) in soil were based on applying ¹⁴C and/or ¹³C labeled substances. Nearly all these studies used uniformly labeled substances, i.e. all C atoms in the molecule were labeled. The underlying premise is that LMWOS transformation involves the whole molecule and it is not possible to distinguish between 1) the flux of the molecule as a whole between pools (i.e. microbial biomass, CO₂, DOM, SOM, etc.) and 2) the splitting of the substance into metabolites and tracing those metabolites within the pools.Based on position-specific¹⁴C labeling, we introduce a new approach for investigating LMWOS transformation in soil: using Na-acetate labeled with ¹⁴C either in the 1st position (carboxyl group, -COOH) or in the 2nd position (methyl group, -CH₃), we evaluated sorption by the soil matrix, decomposition to CO₂, and microbial uptake as related to both C atoms in the acetate. We showed that sorption of acetate occurred as a whole molecule. After microbial uptake, however, the acetate is split, and C from the -COOH group is converted to CO₂ more completely and faster than C from the -CH₃ group. Correspondingly, C from the -CH₃ group of acetate is mainly incorporated into microbial cells, compared to C from the -COOH group. Thus, the rates of C utilization by microorganisms of C from both positions in the acetate were independently calculated. At concentrations of 10 μmol l⁻¹, microbial uptake from soil solution was very fast (half-life time about 3 min) for both C atoms. At concentrations <100 μmol l⁻¹ the oxidation to CO₂ was similar for C atoms of both groups (about 55% of added substance). However, at acetate concentrations >100 μmol l⁻¹, the decomposition to CO₂ for C from -CH₃ decreased more strongly than for C from -COOH.We conclude that the application of position-specifically labeled substances opens new ways to investigate not only the general fluxes, but also transformations of individual C atoms from molecules. This, in turn, allows conclusions to be drawn about the steps of individual transformation processes on the submolecular level and the rates of these processes.     

蚕豆生长及土壤酶活性对低分子量聚乙烯的响应

研究与推广环保型可降解地膜, 是解决当前农田白色污染的重要途径。与普通地膜比较, 可降解地膜降解时间短, 残留物以分子量较低的线性低密度聚乙烯(LLDPE)为主。研究低分子量聚乙烯对作物生长及土壤性质的影响, 对可降解地膜的推广具有重要意义。本研究模拟自然条件, 以添加低分子量聚乙烯(LMWPE)的壤土盆栽种植蚕豆, 并设定不同的LMWPE添加量, 研究全生育期蚕豆的株高、复叶数、结荚数及土壤中脲酶、过氧化物酶、蔗糖酶活性对LMWPE的响应。结果显示: 与不添加LMWPE且种植蚕豆的处理(CK₁)比较, 全生育期中添加LMWPE(分别按覆盖地膜1年、10年、50年、100年折算的添加量)且种植蚕豆处理的蚕豆株高、复叶数及土壤脲酶活性、过氧化物酶活性显著高于CK₁, 且随着LMWPE添加量增加, 土壤酶活性及蚕豆株高和复叶数增幅越显著, 但各处理土壤蔗糖酶活性及蚕豆结荚数差异不显著。蚕豆根系的形成和生长对3种土壤酶活性具有显著影响。3种土壤酶活性与株高、复叶数显著相关, 但与结荚数无显著的相关性。本研究结果表明, 土壤中添加LMWPE对蚕豆生长及3种土壤酶活性有激活作用, 以添加量为2.80 g·kg^-1处理的影响最为明显。     

The contribution of biogas residues to soil organic matter formation and CO2 emissions in an arable soil

The biogas production process generates as side-products biogas residues containing microbial biomass which could contribute to soil organic matter formation or induce CO₂ emissions when applied to arable soil as fertilizer. Using an isotope labelling approach, we labelled the microbial biomass in biogas residues, mainly G⁺ bacteria and methanogenic archaea via KH¹³CO₃, and traced the fate of microbial biomass carbon in soil with an incubation experiment lasting 378 days. Within the first seven days, 40% of the carbon was rapidly mineralized and after that point mineralization continued, reaching 65% by the end of the experiment. Carbon mineralization data with 93% recovery could be fitted to a two-pool degradation model which estimated proportions and degradation rate constants of readily and slowly degrading pools. About 49% of the carbon was in the slowly degrading pool with a half-life of 1.9 years, suggesting mid-term contribution to living and non-living soil organic matter formation. Biogas residues caused a priming effect at the beginning, thus their intensive application should be avoided.     

The impact of a low humus level in arable soils on microbial properties, soil organic matter quality and crop yield

 In arable soils in Schleswig-Holstein (Northwest Germany) nearly 30% of the total organic C (TOC) stored in former times in the soil has been mineralized in the last 20 years. Microbial biomass, enzyme activities and the soil organic matter (SOM) composition were investigated in order to elucidate if a low TOC level affects microbial parameters, SOM quality and crop yield. Microbial biomass C (C_mic) and enzyme activities decreased in soils with a low TOC level compared to soils with a typical TOC level. The decrease in the C_mic/TOC ratio suggested low-level, steady-state microbial activity. The SOM quality changed with respect to an enrichment of initial litter compounds in the top soil layers with a low TOC level. Recent management of the soils had not maintained a desirable level of humic compounds. However, we found no significant decrease in crop yield. We suggest that microbial biomass and dehydrogenase and alkaline phosphatase activities are not necessarily indicators of soil fertility in soils with a high fertilization level without forage production and manure application. Received: 12 December 1997     

长期添加外源有机物料对华北平原不同粒级土壤氮素和氨基糖的影响

华北平原是我国主要的粮食生产基地之一,农民为了追求高产,过量施用化肥的弊端日益凸显。本研究依托中国科学院栾城农业生态系统试验站有机养分循环再利用长期定位试验,开展不同外源有机物料对土壤氮素和氨基糖在不同粒级土壤库中分布的影响研究,为阐释不同农业管理措施下土壤氮素的物理保护机制和生物保护机制提供依据。定位试验设6个处理：无肥无秸秆处理（对照,CK）、单施猪圈肥（M）、单施化肥（NPK）、单施秸秆（SCK）、化肥配施猪圈肥（MNPK）和化肥配施秸秆（SNPK）。通过超声波分散-离心分离得到3种粒径土壤——砂粒级（2 000~53 μm）、粉粒级（53~2 μm）和黏粒级（<2 μm）,分析全土及各粒级土壤中全氮和3种土壤氨基糖（氨基葡萄糖、胞壁酸和氨基半乳糖）的含量及变化;基于这3种土壤氨基糖的稳定性和异源性,以氨基糖作为微生物残留物标识物,了解真菌和细菌残留物的积累和转化,阐释真菌和细菌在养分转化中的作用。结果表明：添加有机物料（秸秆、猪圈肥）明显提升了土壤全氮和氨基糖含量,粒级间土壤氮素和氨基糖含量顺序均为黏粒级>砂粒级>粉粒级。添加有机物料对砂粒级土壤氮素影响最大,长期化肥配施猪圈肥中氮素主要在砂粒级中富集,长期化肥配施秸秆的氮素主要在黏粒级中富集。添加秸秆主要提高了真菌来源的氨基葡萄糖的含量,而添加猪圈肥主要提高了土壤中细菌来源的胞壁酸含量,表明添加不同有机物料可影响土壤微生物的群落结构。从各粒级中氨基葡萄糖/胞壁酸的比值来看,添加不同类型外源有机物料对砂粒级土壤微生物群落结构影响最为明显。由此可见,在长期秸秆还田措施下实施有机粪肥部分替代化肥不仅可以减少化肥用量,还可提升土壤养分含量和微生物多样性,改善土壤质量。     

Long-term fertilization regimes influence FAME profiles of microbial communities in an arable sandy loam soil in Northern China

Soil samples collected from a long-term (19-year) experimental field with seven treatments were analyzed for fatty acids methyl esters (FAMEs) to determine fertilization regime effects on microbial community structure in sandy loam soils. The amounts of FAMEs in bacteria, actinomycetes, and fungi were highest with the two organic manure (OM)-fertilized treatments (OM and 1/2 OMN – half OM plus half mineral fertilizer), lowest with the NK treatment, and fell in the middle levels with three mineral P-fertilized treatments (NPK, NP and PK) and the control with no fertilizer (CK), with the exception of fungi which showed no significant difference among the five treatments without OM fertilization. Principal component analysis of FAME patterns indicated that NPK was not significantly different from CK, but the two manure-containing treatments and the P-deficiency treatment (NK) were significantly different from CK and NPK. Redundancy analysis plot showed that FAME amounts significantly correlated to soil organic C and total N contents, while soil available P and total P contents, which were greatly decreased by the NK treatment, also had positive and substantial effects on soil microbial FAMEs. The results demonstrated the importance of P fertilization as well as organic manure in maintaining soil microbial biomass and impacting community structure.     

Experimental evaluation of methods to quantify dissolved organic nitrogen (DON) and dissolved organic carbon (DOC) in soil

A significant proportion of the total nutrient in soil solution can be bound to organic molecules and these often constitute a major loss from soil to freshwater. Our purpose was to determine whether chemical extractants used for measuring inorganic N could also be used to quantify dissolved organic nitrogen (DON) and carbon (DOC) in soil. In a range of soils, DOC and DON were extracted with either distilled water or 2 M KCl and the amount recovered compared with that present in soil solution recovered by centrifugal-drainage. The recovery of DON and DOC from soil was highly dependent upon the method of extraction. Factors such as soil sampling strategy (number of samples over space and time), sample preparation (sieving and drying), soil storage, extraction temperature, shaking time, and soil-to-extractant volume ratio all significantly affected the amount of DOC and DON extracted from soil. To allow direct comparison between independent studies we therefore propose the introduction of a standardized extraction procedure: Replicate samples of unsieved, field-moist soil extracted as soon as possible after collection with distilled water, 0.5 M K₂SO₄ or 2 M KCl at a 1:5 w/v ratio for 1 h at 20 °C.     

暗棕壤中低分子有机酸的吸附与生物降解研究

低分子有机酸在森林生态系统中普遍存在,在根际和多种成土过程中发挥重要作用,但有机酸发挥作用的能力和效率与其在土壤中的吸附与降解等行为关系密切。本文研究了草酸、柠檬酸和苹果酸等低分子有机酸在森林暗棕壤A1层和B层的吸附及生物降解行为,结果表明,低分子有机酸加入到暗棕壤后迅速被土壤固相颗粒所吸附,其中对草酸的吸附远强于柠檬酸和苹果酸,三种低分子有机酸在暗棕壤A1层的吸附率强于B层;草酸、柠檬酸和苹果酸加入到森林暗棕壤后还能迅速被微生物降解,其中对草酸的降解远低于柠檬酸和苹果酸,暗棕壤对草酸较强的吸附作用可能是其生物降解率较低的主要原因,且三种低分子有机酸在A1层暗棕壤中的生物降解率高于B层。